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FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy
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| Zeitschriftentitel: | Journal of Translational Medicine |
|---|---|
| Personen und Körperschaften: | , , , , , , , , , , |
| In: | Journal of Translational Medicine, 8, 2010, 1 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Springer Science and Business Media LLC
|
| Schlagwörter: |
| author_facet |
Fiammenghi, Laura Ancarani, Valentina Rosales, Tilman Knutson, Jay R Petrini, Massimiliano Granato, Anna Maria Pancisi, Elena Ridolfi, Laura Ridolfi, Ruggero Riccobon, Angela Neyroz, Paolo Fiammenghi, Laura Ancarani, Valentina Rosales, Tilman Knutson, Jay R Petrini, Massimiliano Granato, Anna Maria Pancisi, Elena Ridolfi, Laura Ridolfi, Ruggero Riccobon, Angela Neyroz, Paolo |
|---|---|
| author |
Fiammenghi, Laura Ancarani, Valentina Rosales, Tilman Knutson, Jay R Petrini, Massimiliano Granato, Anna Maria Pancisi, Elena Ridolfi, Laura Ridolfi, Ruggero Riccobon, Angela Neyroz, Paolo |
| spellingShingle |
Fiammenghi, Laura Ancarani, Valentina Rosales, Tilman Knutson, Jay R Petrini, Massimiliano Granato, Anna Maria Pancisi, Elena Ridolfi, Laura Ridolfi, Ruggero Riccobon, Angela Neyroz, Paolo Journal of Translational Medicine FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy General Biochemistry, Genetics and Molecular Biology General Medicine |
| author_sort |
fiammenghi, laura |
| spelling |
Fiammenghi, Laura Ancarani, Valentina Rosales, Tilman Knutson, Jay R Petrini, Massimiliano Granato, Anna Maria Pancisi, Elena Ridolfi, Laura Ridolfi, Ruggero Riccobon, Angela Neyroz, Paolo 1479-5876 Springer Science and Business Media LLC General Biochemistry, Genetics and Molecular Biology General Medicine http://dx.doi.org/10.1186/1479-5876-8-52 <jats:title>Abstract</jats:title> <jats:sec> <jats:title>Background</jats:title> <jats:p>Antigen processing by dendritic cells (DC) exposed to specific stimuli has been well characterized in biological studies. Nonetheless, the question of whether autologous whole tumor lysates (as used in clinical trials) are similarly processed by these cells has not yet been resolved.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>In this study, we examined the transfer of peptides from whole tumor lysates to major histocompatibility complex class II molecules (MHC II) in mature dendritic cells (mDC) from a patient with advanced melanoma. Tumor antigenic peptides-MHC II proximity was revealed by Förster Resonance Energy Transfer (FRET) measurements, which effectively extends the application of fluorescence microscopy to the molecular level (<100Å). Tumor lysates were labelled with Alexa-488, as the donor, and mDC MHC II HLA-DR molecules were labelled with Alexa-546-conjugated IgG, as the acceptor.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>We detected significant energy transfer between donor and acceptor-labelled antibodies against HLA-DR at the membrane surface of mDC. FRET data indicated that fluorescent peptide-loaded MHC II molecules start to accumulate on mDC membranes at 16 hr from the maturation stimulus, steeply increasing at 22 hr with sustained higher FRET detected up to 46 hr.</jats:p> </jats:sec> <jats:sec> <jats:title>Conclusions</jats:title> <jats:p>The results obtained imply that the patient mDC correctly processed the tumor specific antigens and their display on the mDC surface may be effective for several days. These observations support the rationale for immunogenic efficacy of autologous tumor lysates.</jats:p> </jats:sec> FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy Journal of Translational Medicine |
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Springer Science and Business Media LLC, 2010 |
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2010 |
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| title |
FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_unstemmed |
FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_full |
FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_fullStr |
FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_full_unstemmed |
FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_short |
FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_sort |
fret microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| topic |
General Biochemistry, Genetics and Molecular Biology General Medicine |
| url |
http://dx.doi.org/10.1186/1479-5876-8-52 |
| publishDate |
2010 |
| physical |
|
| description |
<jats:title>Abstract</jats:title>
<jats:sec>
<jats:title>Background</jats:title>
<jats:p>Antigen processing by dendritic cells (DC) exposed to specific stimuli has been well characterized in biological studies. Nonetheless, the question of whether autologous whole tumor lysates (as used in clinical trials) are similarly processed by these cells has not yet been resolved.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Methods</jats:title>
<jats:p>In this study, we examined the transfer of peptides from whole tumor lysates to major histocompatibility complex class II molecules (MHC II) in mature dendritic cells (mDC) from a patient with advanced melanoma. Tumor antigenic peptides-MHC II proximity was revealed by Förster Resonance Energy Transfer (FRET) measurements, which effectively extends the application of fluorescence microscopy to the molecular level (<100Å). Tumor lysates were labelled with Alexa-488, as the donor, and mDC MHC II HLA-DR molecules were labelled with Alexa-546-conjugated IgG, as the acceptor.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Results</jats:title>
<jats:p>We detected significant energy transfer between donor and acceptor-labelled antibodies against HLA-DR at the membrane surface of mDC. FRET data indicated that fluorescent peptide-loaded MHC II molecules start to accumulate on mDC membranes at 16 hr from the maturation stimulus, steeply increasing at 22 hr with sustained higher FRET detected up to 46 hr.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Conclusions</jats:title>
<jats:p>The results obtained imply that the patient mDC correctly processed the tumor specific antigens and their display on the mDC surface may be effective for several days. These observations support the rationale for immunogenic efficacy of autologous tumor lysates.</jats:p>
</jats:sec> |
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| author | Fiammenghi, Laura, Ancarani, Valentina, Rosales, Tilman, Knutson, Jay R, Petrini, Massimiliano, Granato, Anna Maria, Pancisi, Elena, Ridolfi, Laura, Ridolfi, Ruggero, Riccobon, Angela, Neyroz, Paolo |
| author_facet | Fiammenghi, Laura, Ancarani, Valentina, Rosales, Tilman, Knutson, Jay R, Petrini, Massimiliano, Granato, Anna Maria, Pancisi, Elena, Ridolfi, Laura, Ridolfi, Ruggero, Riccobon, Angela, Neyroz, Paolo, Fiammenghi, Laura, Ancarani, Valentina, Rosales, Tilman, Knutson, Jay R, Petrini, Massimiliano, Granato, Anna Maria, Pancisi, Elena, Ridolfi, Laura, Ridolfi, Ruggero, Riccobon, Angela, Neyroz, Paolo |
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| description | <jats:title>Abstract</jats:title> <jats:sec> <jats:title>Background</jats:title> <jats:p>Antigen processing by dendritic cells (DC) exposed to specific stimuli has been well characterized in biological studies. Nonetheless, the question of whether autologous whole tumor lysates (as used in clinical trials) are similarly processed by these cells has not yet been resolved.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>In this study, we examined the transfer of peptides from whole tumor lysates to major histocompatibility complex class II molecules (MHC II) in mature dendritic cells (mDC) from a patient with advanced melanoma. Tumor antigenic peptides-MHC II proximity was revealed by Förster Resonance Energy Transfer (FRET) measurements, which effectively extends the application of fluorescence microscopy to the molecular level (<100Å). Tumor lysates were labelled with Alexa-488, as the donor, and mDC MHC II HLA-DR molecules were labelled with Alexa-546-conjugated IgG, as the acceptor.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>We detected significant energy transfer between donor and acceptor-labelled antibodies against HLA-DR at the membrane surface of mDC. FRET data indicated that fluorescent peptide-loaded MHC II molecules start to accumulate on mDC membranes at 16 hr from the maturation stimulus, steeply increasing at 22 hr with sustained higher FRET detected up to 46 hr.</jats:p> </jats:sec> <jats:sec> <jats:title>Conclusions</jats:title> <jats:p>The results obtained imply that the patient mDC correctly processed the tumor specific antigens and their display on the mDC surface may be effective for several days. These observations support the rationale for immunogenic efficacy of autologous tumor lysates.</jats:p> </jats:sec> |
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| imprint | Springer Science and Business Media LLC, 2010 |
| imprint_str_mv | Springer Science and Business Media LLC, 2010 |
| institution | DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229 |
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| spelling | Fiammenghi, Laura Ancarani, Valentina Rosales, Tilman Knutson, Jay R Petrini, Massimiliano Granato, Anna Maria Pancisi, Elena Ridolfi, Laura Ridolfi, Ruggero Riccobon, Angela Neyroz, Paolo 1479-5876 Springer Science and Business Media LLC General Biochemistry, Genetics and Molecular Biology General Medicine http://dx.doi.org/10.1186/1479-5876-8-52 <jats:title>Abstract</jats:title> <jats:sec> <jats:title>Background</jats:title> <jats:p>Antigen processing by dendritic cells (DC) exposed to specific stimuli has been well characterized in biological studies. Nonetheless, the question of whether autologous whole tumor lysates (as used in clinical trials) are similarly processed by these cells has not yet been resolved.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>In this study, we examined the transfer of peptides from whole tumor lysates to major histocompatibility complex class II molecules (MHC II) in mature dendritic cells (mDC) from a patient with advanced melanoma. Tumor antigenic peptides-MHC II proximity was revealed by Förster Resonance Energy Transfer (FRET) measurements, which effectively extends the application of fluorescence microscopy to the molecular level (<100Å). Tumor lysates were labelled with Alexa-488, as the donor, and mDC MHC II HLA-DR molecules were labelled with Alexa-546-conjugated IgG, as the acceptor.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>We detected significant energy transfer between donor and acceptor-labelled antibodies against HLA-DR at the membrane surface of mDC. FRET data indicated that fluorescent peptide-loaded MHC II molecules start to accumulate on mDC membranes at 16 hr from the maturation stimulus, steeply increasing at 22 hr with sustained higher FRET detected up to 46 hr.</jats:p> </jats:sec> <jats:sec> <jats:title>Conclusions</jats:title> <jats:p>The results obtained imply that the patient mDC correctly processed the tumor specific antigens and their display on the mDC surface may be effective for several days. These observations support the rationale for immunogenic efficacy of autologous tumor lysates.</jats:p> </jats:sec> FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy Journal of Translational Medicine |
| spellingShingle | Fiammenghi, Laura, Ancarani, Valentina, Rosales, Tilman, Knutson, Jay R, Petrini, Massimiliano, Granato, Anna Maria, Pancisi, Elena, Ridolfi, Laura, Ridolfi, Ruggero, Riccobon, Angela, Neyroz, Paolo, Journal of Translational Medicine, FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy, General Biochemistry, Genetics and Molecular Biology, General Medicine |
| title | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_full | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_fullStr | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_full_unstemmed | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_short | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_sort | fret microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| title_unstemmed | FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy |
| topic | General Biochemistry, Genetics and Molecular Biology, General Medicine |
| url | http://dx.doi.org/10.1186/1479-5876-8-52 |